陕西省2018年布鲁氏菌种型鉴定方法的比较分析

Comparative analysis of identification methods of Brucella strains in Shaanxi Province in 2018

目的对陕西省2018年分离的疑似布鲁氏菌进行种型鉴定,为布病防治提供科学依据。方法应用形态学鉴定、BCSP31、AMOS、16SrDNA-PCR及生化鉴定方法,对2018年全省收集的30株疑似布鲁氏菌进行鉴定。结果BCSP31-PCR检测结果中,1、33号菌株为阴性,12、19、20、23、29、30、31号菌株疑似,其余均为阳性;AMOS-PCR扩增结果显示,BCSP31-PCR阴性及疑似的菌株均为阴性,其余均出现731 bp左右的目的条带,确定为羊种菌;16SrDNA-PCR结果与AMOS-PCR一致,AMOS-PCR中未出现条带的9株菌均显示为其他菌,其余菌株与布鲁氏菌的同源性99%;生化鉴定确定4号为羊1型,3、11、32号为羊种变异株,其余均为羊3型。结论 2018年分离获得的30株菌经鉴定有9株排除是布鲁氏菌,在实际工作中,尤其是遇到疑似菌株时应综合应用,多种方法相互印证,以获得更准确的鉴定结果。

Objective To identify the species of suspected Brucella isolated in Shaanxi Province in 2018,so as to provide scientific basis for brucellosis prevention and control.Methods 30 strains of suspected Brucella collected in the province in 2018 were identified by morphological identification,BCSP31,AMOS,16 SrDNA-PCR and biochemical identification methods.Results The BCSP31-PCR results showed that strains 1 and 33 were negative,and strains 12,19,20,23,29,30 and 31 were suspected,and the rest were positive.AMOS-PCR amplification results showed that all the BCSP31-PCR negative and suspected strains were negative,and all the others showed target bands of about 731 bp,which were identified as Bmelitensis strains.The results of 16 SrDNA-PCR were consistent with that of AMOS-PCR.All the 9 strains without bands in AMOS-PCR were identified as other strains,and the homology of the remaining strains was 99%with Brucella.Biochemical identification confirmed that the strain of No.4 was melitensisⅠ,and No.3,No.11 and No.32 were melitensis variant strains,and the rest were melitensisⅢ.Conclusion 9 of the 30 strains isolated and obtained in 2018 have been identified as Brucella.In practical work,especially when encountering suspected strains,a comprehensive application should be made to verify each other with multiple methods so as to obtain more accurate identification results.